With the development of microscopic biology, requirements for microscopic imaging technology have been increased. Acquiring a wide-field and high-resolution microscopic image in real time is a precondition for microscopic biology research. For a cancer research, for example, a high-resolution image is needed for observing a propagation step of cancer cells, while a wide field image is required for observing a migration of the cancer cells. In addition, acquiring the microscopic image at high speed and in real time is needed for observing a fast transmission of a signal via a neuron.
An image observed via an ocular of a microscope may be converted to a wide-field surface intermediary image via a series of objective lens groups, and sub-field-of-view high-resolution images can be obtained by performing a divided field collection on the surface intermediary image by the image sensor array. In order to gather the sub-field-of-view high-resolution images together to acquire a wide-field high-resolution image, a calibration on the image sensor array in a microscopic field is necessary.
Compared to a conventional imaging condition in the macroscopic scenario, it is rather a challenge to calibrate the image sensor array in the microscopic field. Firstly, due to a limitation of a shallow depth of focus of microscope objective lens, three-dimensional information of the calibration object is unable to be acquired by changing a location thereof. Additionally, it is difficult to perform the calibration step by step using a system includes multistage objective lens and cambered intermediate image. Therefore, a method for calibrating an image sensor array applied in a microscopic imaging system is needed, for satisfying a requirement of obtaining a wide-field high-resolution and real-time microscopic image.